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Published on: June 25, 2020
Rapid detection of squash leaf curl virus by loop-mediated isothermal amplification
Cheng-Ping Kuan1, Min-Tze Wu, Yi-Lin Lu
1Division of Biotechnology, Taiwan Agricultural Research Institute, No. 189 Chung Cheng Rd, Wufeng, Taichung 41362, Taiwan. pcr123@tari.gov.tw
Journal of Virological Methods
|July 7, 2010
Summary
A new loop-mediated isothermal amplification (LAMP) assay efficiently detects squash leaf curl virus (SLCV) in plants. This rapid LAMP method is more sensitive and simpler than traditional PCR for diagnosing viral infections in squash and melon.
Area of Science:
- Plant Pathology
- Molecular Biology
- Virology
Background:
- Squash leaf curl virus (SLCV) causes significant damage to cucurbitaceous crops like squash and melon.
- Accurate and rapid detection of SLCV is crucial for effective disease management.
Purpose of the Study:
- To develop a simple, efficient, and sensitive diagnostic system for SLCV detection.
- To compare the efficacy of the developed Loop-mediated Isothermal Amplification (LAMP) assay with Polymerase Chain Reaction (PCR).
Main Methods:
- A set of four primers targeting SLCV was designed for LAMP assay.
- The LAMP assay was performed under isothermal conditions at 65°C for 30-60 minutes.
- LAMP products were visualized using SYBR Safe DNA gel stain.
Main Results:
- The LAMP assay successfully detected SLCV in infected squash and melon plants.
- LAMP demonstrated a 10-fold higher sensitivity than PCR for SLCV detection in diseased plants.
- No cross-reactivity was observed in healthy plant tissues with either LAMP or PCR.
Conclusions:
- The developed LAMP assay is a rapid, simple, accurate, and sensitive method for SLCV detection.
- LAMP offers a more practical alternative to PCR for diagnosing SLCV infections in cucurbitaceous plants.
